Multiplexed 3D Cellular Super-Resolution Imaging with DNA-PAINT and Exchange-PAINT

By R. Jungmann, Maier S. Avendaño, Johannes B. Woehrstein, Mingjie Dai, William M. Shih, Pengcheng Yin. Nature Methods, 2014

1,007 citations recorded by the source corpus.

Super-resolution fluorescence microscopy is a powerful tool for biological research, but obtaining multiplexed images for a large number of distinct target species remains challenging. Here we use the transient binding of short fluorescently labeled oligonucleotides (DNA-PAINT, a variation of point accumulation for imaging in nanoscale topography) for simple and easy-to-implement multiplexed super-resolution imaging that achieves sub-10-nm spatial resolution in vitro on synthetic DNA structures. We also report a multiplexing approach (Exchange-PAINT) that allows sequential imaging of multiple targets using only a single dye and a single laser source. We experimentally demonstrate ten-color super-resolution imaging in vitro on synthetic DNA structures as well as four-color two-dimensional (2D) imaging and three-color 3D imaging of proteins in fixed cells.

Subfields: Biomedical Engineering, Biophysics, Structural Biology

View Paper: https://www.semanticscholar.org/paper/bc59ec243ed46360e0dac63b439cda63a8c89106

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In Biomedical Engineering

1. Florian Jug
2. Yoav Shechtman
3. Loic A. Royer
4. M. Heilemann
5. Elias Nehme

In Biophysics

1. Florian Jug
2. Alexander Krull
3. Tim-Oliver Buchholz
4. Loic A. Royer
5. M. Heilemann
6. Elias Nehme
7. D. Bull
8. Nantheera Anantrasirichai

In Structural Biology

1. Florian Jug
2. Yoav Shechtman
3. M. Heilemann
4. Elias Nehme

Citations in the corpus

• Deep learning massively accelerates super-resolution localization microscopy (2018, background): “Adaptation of ANNA-PALM to 3D 4,45 and multicolor 46,47 localization microscopy should be relatively straightforward.”
• The Beauty and Utility of DNA Origami (2017, methodology): “Reprinted by permission from Macmillan Publishers Ltd: Nature Methods (Jungmann et al.(77)), copyright 2014.”
• Democratising deep learning for microscopy with ZeroCostDL4Mic (2021, methodology): “In all, 2 nM P1-ATTO655 (8 nt duplex, Euro fi ns) in PBS pH 8.2 + 500 mM NaCl was added to the sample, resulting in a labelling density signi fi cantly higher than used for conventional DNA-PAINT imaging.”